Molecular Detection by PCR

Molecular detection of Chlamydophila pneumoniae by PCR.

Bronchioalveolar lavage (BAL) or lung biopsy. We may also perform testing on nasopharyngeal aspirate (NPA), sputum, and cerebrospinal fluid (CSF) however please note that these specimens are not ideal for testing. Dry swabs are not acceptable for testing; any swabs sent must be supplied in appropriate storage medium. At least 1 mL of fluid sample is required for testing, and it is preferable if all samples are supplied in screw-capped tubes made of freeze-thaw and shatter-resistant plastic.

Please follow standard aseptic sampling methods for collecting specimens and ensure any tissue samples/swabs are in appropriate storage medium.

No further processing at the sending lab is required once specimens are collected according to the above instructions. All specimens should be stored at refrigeration temperature (2 – 8°C) or frozen after collection. Ensure that specimens are held at the appropriate temperature when shipped to the NML (e.g. frozen specimens shipped with dry ice, refrigerated specimens with cold packs).

Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.

Symptoms of respiratory infection (e.g. bronchitis, atypical pneumonia) are indicative of possible C. pneumoniae infection. However it has also been associated with other disorders such as coronary artery disease, asthma, and sarcoidosis¹

Completed Special Bacteriology requisition form detailing all patient information and relevant clinical information. If possible, attach lab results that have already been done at local or provincial laboratories. Documentation must include clinical background leading to suspicion of C. pneumoniae infection in a patient.

If a test performed by the submitting lab produced a positive result for C. pneumoniae, it is also helpful to indicate this in the documentation including what type of test was performed.

DNA from all C. pneumoniae samples received at the NML for PCR testing is initially extracted using a commercially-available kit. Results are based upon a real-time PCR assay targeting a highly conserved 16S ribosomal gene sequence within C. pneumonia. PCR to detect the human b-globin gene to test for DNA quality and integrity is also performed and must be positive.

12 calendar days. Please note that during times when large numbers of samples are received or if tests must be repeated the turnaround time may be longer.

1. Hardick J, Maldeis N, Throdore M, Wood BJ, Yang S, Lin S, Quinn T and Gaydos C. Real-Time PCR for Chlamydia pneumoniae utilizing the Roche Lightcycler and a 16SrRNA gene target. (2004), Journal of Molecular Diagnostics 6(2):132-136.